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Vibrio Fluvialis: an Emerging Human Pathogen

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Vibrio Fluvialis: an Emerging Human Pathogen REVIEW ARTICLE published: 07 March 2014 doi: 10.3389/fmicb.2014.00091 Vibrio fluvialis: an emerging human pathogen Thandavarayan Ramamurthy 1*, Goutam Chowdhury 1, Gururaja P.Pazhani 1 and Sumio Shinoda 2 1 National Institute of Cholera and Enteric Diseases, Kolkata, India 2 National Institute of Cholera and Enteric Diseases, Collaborative Research Center of Okayama University for Infectious Diseases in India, Kolkata, India Edited by: Vibrio fluvialis is a pathogen commonly found in coastal environs. Considering recent Rita R. Colwell, University of increase in numbers of diarrheal outbreaks and sporadic extraintestinal cases, V.fluvialis has Maryland, USA been considered as an emerging pathogen. Though this pathogen can be easily isolated Reviewed by: by existing culture methods, its identification is still a challenging problem due to close Carlos R. Osorio, University of Santiago de Compostela, Spain phenotypic resemblance either with Vibrio cholerae or Aeromonas spp. However, using Brian Austin, University of Stirling, UK molecular tools, it is easy to identify V. fluvialis from clinical and different environmental *Correspondence: samples. Many putative virulence factors have been reported, but its mechanisms of Thandavarayan Ramamurthy, National pathogenesis and survival fitness in the environment are yet to be explored. This chapter Institute of Cholera and Enteric covers some of the major discoveries that have been made to understand the importance Diseases, P-33, CIT Road, Scheme-XM, Beliaghata, of V. fluvialis. Kolkata-700010, India Keywords:V. fluvialis, diarrhea, virulence factors, antimicrobial resistance, molecular typing e-mail: [email protected] INTRODUCTION importance of V. fluvialis (Chowdhury et al., 2012; Liang et al., Vibrio fluvialis is a halophilic Gram-negative bacterium, which 2013). has a curved cell morphology and polar flagella for motility. The important biochemical features of this organism include IDENTIFICATION AND TAXONOMY conversion of nitrate to nitrite, do not cleave L-lysine or Thiosulfate-citrate-bile salts-sucrose agar (TCBS) has been con- ornithine, activate arginine dihydrolase, produce indole but ventionally used as a selective medium for the isolation of clinically not acetoin, ferment sucrose, D-mannitol, L-arabinose, maltose, important vibrios. The colony morphology of V. fluvialis in this trehalose, D-galactose, and D-galacturonate. Most of the vib- medium remains indistinguishable from V. cholerae, i.e., it grows rios, including V. fluvialis occur widely in the aquatic milieu, as sucrose fermenting yellow color colonies after direct plating of mostly in the seas, estuaries and brackish waters. Even though clinical specimens or after enrichment in alkaline peptone water more than 100 spices have been reported in the Genus Vib- (pH 8.0). After preliminary screening in the TCBS, a battery rio (http://www.bacterio.net/uw/vibrio.html), about 13 of them of biochemical testes is essential for the species-specific identi- have been reported to cause several human diseases. Among fication of V. fluvialis. Minimal biochemical tests such as lysine the pathogenic vibrios, V. alginolyticus, V. cholerae, V. costicola, decarboxylase, ornithine decarboxylase, arginine dihydrolase, and V. mimicus, V. cincinnatiensis, V. hollisae, V. furnissii, V. para- L-arabinose are mandatory for the identification of V. fluvialis. haemolyticus, V. vulnificus, V. carchariae (a junior synonym of Without these minimal tests, the identification is incomplete and V. harveyi) and V. metschnikovii are clinically important as they the isolate will be improperly classified as V. cholerae or Aeromonas cause different types of vibriosis. One of the Vibrio spp., V. spp. In most resource-poor countries, these tests are not method- damselae has now been renamed as “Photobacterium damselae ically performed, which may lead to labeling of V. fluvialis as subsp. damselae.” The toxigenic V. cholerae, V. parahaemolyti- V. cholerae. Considering such situation, there is a high possibil- cus and V. vulnificus are associated with well-known cholera ity that the V. fluvialis could be reported as V. cholerae non-O1, and diarrhea and extraintestinal infections, respectively. Preva- non-O139 or non-agglutinable vibrios (NAGs). It is worth to lence of V. cholerae in developing countries is mostly related mention here that V. cholerae O1 and O139 serogroups can to the breakdown of sanitary conditions and/or due to scarcity be easily confirmed by slide agglutination with corresponding of drinking water. On the other hand, infections caused by antiserum. V. parahaemolyticus and other vibrios denote contamination For the identification of V. fluvialis and other vibrios, rapid of seafood in many countries, irrespective of their economic identification kits must be used with caution as they need conditions. additional tests for the final confirmation. While testing the com- V. fluvialis is one of the emerging foodborne pathogens all over mercially available identification kits, V.fluvialis remain as a major the world. The distribution of virulence factors and molecular epi- challenge with API 20E and Vitek GNI+ systems (Israil et al., 2003; demiological features of this pathogen remain mostly unknown. O’Hara et al., 2003). Biochemically, V. furnissii expresses fibrin Among the foodborne infections in the United States, there and mucin hydrolysis but no phosphate or esculin hydrolysis, for has been a considerable increase (43%) in the Vibrio-mediated which V. fluvialis varied. V. fluvialis, V. furnissii, and V. mimicus are infections till 2012 compared with the rates reported during distinctive from V. cholerae, as the later exhibit strong mannose- 2006–2008 (Centers for Disease Control and Prevention (CDC), sensitive hemagglutination. These test results may have a strong 2013). Several recent publications indicate the epidemiological influence in the confirmation of strains. www.frontiersin.org March 2014 | Volume 5 | Article 91 | 1 Ramamurthy et al. Emerging Vibrio fluvialis Molecular tools such as PCR are useful in the identification for the identification of V. fluvialis and with a set of phages, of many uncommon vibrios and most of these assays are com- the diagnostic probability of human isolates was more than parable to the conventional identification methods. The sequence 84%. At least in one study, the importance of phage-typing of amplified 16S–23S intergenic spacers (IGSs) has demonstrated of V. fluvialis has been demonstrated using six specific bac- 37 ribosomal RNA (rrn) operons representing seven different IGS teriophages with 73% typability (Suthienkul, 1993). However, types in different Vibrio spp. with IGS(0), IGS(IA), and IGS(Glu) availability of these bacteriophages makes this assay technique less as major ones. The sequence difference in these IGS types was popular. used to design species-specific primers for PCR for V. fluvialis and other vibrios (Lee et al., 2002). In some of the reports, a uni- PHENOTYPIC AND GENETIC CHARACTERISTICS OF versal primer PCR that covers conserved regions of bacterial 16S V. fluvialis rRNA genes followed by denaturing gradient gel electrophoresis Based on the somatic antigen variation, several serotypes of (DGGE) was found to be useful in the identification of V. flu- V. fluvialis have been identified. Though Shimada et al. (1999) vialis either as axenic bacteria or mixed with other pathogens identified more than 50 somatic antigens, the serological based (Ji et al., 2004). typing of V. fluvialis remains non-customary. V. fluvialis strains Initially, V. furnissii was taxonomically assigned with V. fluvialis belonging to serogroup O19 possessed the C (Inaba) antigen of and named as aerogenic biogroup of V. fluvialis.BasedonDNA V. cholerae O1, but not the B (Ogawa) or A (common) anti- relatedness and several biochemical tests, V. furnissii has been sep- gens (Shimada et al., 1987; Kondo et al., 2000). In the crossed arated as a new species (Lee et al., 1981; Brenner et al., 1983). In the immuno-electrophoresis, antibodies against the oral cholera vac- phylogenetic analysis with several housekeeping genes, V. furnissii cines containing killed whole cells (WC) of V. cholerae O1 Inaba and V. fluvialis have been linked as close species. The nucleotide El Tor reacted with a few strains of V. fluvialis (Ciznãr et al., 1989). comparison of 16S-rRNA, recA, and toxR sequences showed that Presence of shared WC antigens indicates that the oral cholera V. furnissii and V. fluvialis had 100% similarity. The gene toxR of V. vaccine could stimulate immunity effectively against other vib- fluvialis had 84% similarity with V.harveyi (Franco and Hedreyda, rios also. It is known that the antigenic nature of flagella of 2006). With the gyrB, V. cholerae, V. mimicus, V. furnissii, and V. vibrios is highly homologous. Tassin et al. (1983) and Shinoda fluvialis shared 93% sequence similarity. et al. (1984) demonstrated independently that anti-L-flagella anti- Toxigenic vibrios have a homolog of the toxRS operon, which sera of V. fluvialis did not agglutinate other Vibrio species in the regulates the virulence expression. The gene toxR encodes a tran- H-agglutination tests. Further studies placed V.fluvialis and V.fur- scriptional activation domain (TAD), a transmembrane domain nissii in the same lateral flagellar serogroup-HL8 (Shinoda et al., (TMD), and a periplasmic domain (PD). Among the vibrios, 1992). However, in practice, serotyping based on H-flagella is also there is essentially no homology within the region between TAD not in use. and TMD. Hence, this region has been used
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